Study On The Preparation Of Opuntia Monacantha Injection And Its Fingerprint

In the study, Opuntia monacantha,which was widely used in the treatment of animal diseases in recent years,was selected as the object.We adopted three technicss to study its injection preparation process preliminarily.In order to obtain the optimum technics, stability test,irritative test,hemolysis test,alternative acute toxicity test and verifying relevant materials of injection test were carried out and their results were analysed. On this basis, the chromatographic fingerprints of Opuntia Monacantha, its intermediate and injection were studied by HPLC.This provided a method to control the quality of injection.1 Wild Opuntia monacantha,which was collected from Sichuan Province,was used to develop the injection and the content of total flavone was the index.We adopted the traditional water extraction and alcohol precipitation Technics,the ultrasonic assisted extraction Technics and the macroporous resin Technics in the development process.2 To verify the Opuntia monacantha Injection produced by above three kinds of Technicss,extensive tests,such as stability,irritative,hemolysis and verifying relevant materials of injection,were carried out.Results indicated that the macroporous resin Technics was better than the other two kinds of Technicss.Consequently,the preparation process of the Opuntia monacantha Injection are as follows:the extracting solution which has about 0.72mg-mL-1 total flavone is obtained by ultrasonic assisted extraction,its pH value is 7.0,the feed rate is 1BV·h-1 (BV standing for bed volume)and the total sample volume is 3BV. After adsorption completely, the adsorbate-laden column is washed with distilled water untill the effluent don't contain sugar,and then desorbed with aqueous solution of 70% alcohol,the eluting solvent flowrate is 2BV-h-1 and the total volumn is 3BV.The eluent is collected and concentrated in the rotary evaporation apparatus till there isn't alcohol, then add appropriate amount of distilled water into it,place for 24h and filter it.Adding Tween-80 and EDTA into the filtrate,make sure their content to be 0.25% and 0.01% respectively.At last,adding water make 1mL injection containing 1g raw materials,and regulate pH value to 7.0, filter it with millipore membrane, pour it in ampoules and block the ampoules,then sterilize them by flowing steam for 30min.3 According to the papers of State Food and Drug Administration-the technological prescribe of the research on chinese medicinal injections'Fingerprint Spectrum,the fingerprints of Opuntia Monacantha, its intermediate and injection were studied by HPLC.The chromatographic condition are as follows:Agilent TC-C18(2) analytical column (250mm×4.6mm,5μm);mobile phase is methanol(A)-2% acetic acid aqueous(B),gradient eluent,0-5min,5%A,5-70min,5%-40%A,70-80min,40%-50%A,80-90min,50%-55%A;the flow rate:1mL-min-1sample volume:10μL;column tempreture:25℃;wave length:340nm. The Operating Standard of Similarity Evaluation System for Chromatographic Fingerprint of Chinese Materia Medica (Version 2004A) was used to analyze. So the HPLC reference fingerprints of raw material of Opuntia Monacantha,injection intermediate and injection were established and the similarities of the raw material of Opuntia Monacantha,injection intermediate and injection were more than 0.90 respectively.It is necessary for quality control of Opuntia Monacantha injection.